LOCUS       .                       7492 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        183..211
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
     CDS             261..326
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     CDS             339..3410
                     /label="lacZ"
                     /note="β-galactosidase"
     promoter        3795..3899
                     /label="AmpR promoter"
     CDS             3900..4757
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      4931..5519
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5763..5840
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5841..6920
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    6936..6957
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6972..7002
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     CDS             7046..7219
                     /label="lacZα"
                     /note="LacZα fragment of β-galactosidase"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg gactacaaag accatgacgg tgattataaa gatcatgaca
      301 tcgactacaa ggatgacgat gacaagggtt caggatccat gaccatgatt acggattcac
      361 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
      421 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
      481 cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag
      541 aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc
      601 cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca
      661 ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat
      721 ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgtta
      781 actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag gacagtcgtt
      841 tgccgtctga atttgacctg agcgcatttt tacgcgccgg agaaaaccgc ctcgcggtga
      901 tggtgctgcg ctggagtgac ggcagttatc tggaagatca ggatatgtgg cggatgagcg
      961 gcattttccg tgacgtctcg ttgctgcata aaccgactac acaaatcagc gatttccatg
     1021 ttgccactcg ctttaatgat gatttcagcc gcgctgtact ggaggctgaa gttcagatgt
     1081 gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt atggcagggt gaaacgcagg
     1141 tcgccagcgg caccgcgcct ttcggcggtg aaattatcga tgagcgtggt ggttatgccg
     1201 atcgcgtcac actacgtctg aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga
     1261 atctctatcg tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt gaagcagaag
     1321 cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca
     1381 agccgttgct gattcgaggc gttaaccgtc acgagcatca tcctctgcat ggtcaggtca
     1441 tggatgagca gacgatggtg caggatatcc tgctgatgaa gcagaacaac tttaacgccg
     1501 tgcgctgttc gcattatccg aaccatccgc tgtggtacac gctgtgcgac cgctacggcc
     1561 tgtatgtggt ggatgaagcc aatattgaaa cccacggcat ggtgccaatg aatcgtctga
     1621 ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg gtgcagcgcg
     1681 atcgtaatca cccgagtgtg atcatctggt cgctggggaa tgaatcaggc cacggcgcta
     1741 atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg gtgcagtatg
     1801 aaggcggcgg agccgacacc acggccaccg atattatttg cccgatgtac gcgcgcgtgg
     1861 atgaagacca gcccttcccg gctgtgccga aatggtccat caaaaaatgg ctttcgctac
     1921 ctggagagac gcgcccgctg atcctttgcg aatacgccca cgcgatgggt aacagtcttg
     1981 gcggtttcgc taaatactgg caggcgtttc gtcagtatcc ccgtttacag ggcggcttcg
     2041 tctgggactg ggtggatcag tcgctgatta aatatgatga aaacggcaac ccgtggtcgg
     2101 cttacggcgg tgattttggc gatacgccga acgatcgcca gttctgtatg aacggtctgg
     2161 tctttgccga ccgcacgccg catccagcgc tgacggaagc aaaacaccag cagcagtttt
     2221 tccagttccg tttatccggg caaaccatcg aagtgaccag cgaatacctg ttccgtcata
     2281 gcgataacga gctcctgcac tggatggtgg cgctggatgg taagccgctg gcaagcggtg
     2341 aagtgcctct ggatgtcgct ccacaaggta aacagttgat tgaactgcct gaactaccgc
     2401 agccggagag cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg aacgcgaccg
     2461 catggtcaga agccgggcac atcagcgcct ggcagcagtg gcgtctggcg gaaaacctca
     2521 gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct gaccaccagc gaaatggatt
     2581 tttgcatcga gctgggtaat aagcgttggc aatttaaccg ccagtcaggc tttctttcac
     2641 agatgtggat tggcgataaa aaacaactgc tgacgccgct gcgcgatcag ttcacccgtg
     2701 caccgctgga taacgacatt ggcgtaagtg aagcgacccg cattgaccct aacgcctggg
     2761 tcgaacgctg gaaggcggcg ggccattacc aggccgaagc agcgttgttg cagtgcacgg
     2821 cagatacact tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag catcagggga
     2881 aaaccttatt tatcagccgg aaaacctacc ggattgatgg tagtggtcaa atggcgatta
     2941 ccgttgatgt tgaagtggcg agcgatacac cgcatccggc gcggattggc ctgaactgcc
     3001 agctggcgca ggtagcagag cgggtaaact ggctcggatt agggccgcaa gaaaactatc
     3061 ccgaccgcct tactgccgcc tgttttgacc gctgggatct gccattgtca gacatgtata
     3121 ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa ttgaattatg
     3181 gcccacacca gtggcgcggc gacttccagt tcaacatcag ccgctacagt caacagcaac
     3241 tgatggaaac cagccatcgc catctgctgc acgcggaaga aggcacatgg ctgaatatcg
     3301 acggtttcca tatggggatt ggtggcgacg actcctggag cccgtcagta tcggcggaat
     3361 tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa ggatctggct
     3421 cattctctac cccggtttgg atctctcagg ctcagggtat ccgtgctggt ccgtgactcg
     3481 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     3541 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3601 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     3661 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     3721 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3901 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3961 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4021 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4081 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4141 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4201 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4261 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4321 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4381 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4441 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4501 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4561 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4621 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4681 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4741 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4801 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4861 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4921 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4981 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5041 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5101 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5161 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5221 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5281 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5341 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5401 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5461 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5521 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5581 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5641 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5701 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     5761 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     5821 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     5881 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     5941 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     6001 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     6061 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     6121 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     6181 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     6241 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     6301 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     6361 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     6421 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     6481 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     6541 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     6601 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     6661 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     6721 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     6781 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     6841 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6901 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6961 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7021 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     7081 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     7141 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     7201 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     7261 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     7321 gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta cggtcaatcc
     7381 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     7441 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa tt
//
